1. BIOSAFETY
LEVELS
Level 1,2,3,4

2. HISTORY
The first prototype Class III (maximum
containment) biosafety cabinet was
fashioned in 1943 by Hubert Kaempf Jr
April 18, 1955 14 Representatives met at camp
Detrick in Frederick, Maryland.
Purpose: To share knowledge and experiences
regarding bio-safety, chemical, radiological, and
industrial safety issues that were common at the
three principal laboratories of the U.S. Army
1st Biological Safety Conference.
CDC, 1964

3. HISTORY
Center for Disease Control (CDC) specifies the
biosafety levels for the U.S.
BSL1-BSL4
The standard practices pertain to all levels
Bio-safety Level- Level of the bio-containment
precautions required to isolate dangerous
biological agents in an enclosed facility.

4. BIOHAZARD SYMBOL
Charles Baldwin at
National Cancer Institute
at NIH.
Symbol to be
“memorable but
meaningless” so it could
be learned.
Blaze orange – most
visible under harsh
conditions

5. BIOSAFETY LEVEL
1

6. BSL 1
BIOSAFETY 1 is suitable for work involving
well-characterized agents not known to cause
disease in healthy adult humans, and of minimal
potential hazard to laboratory personnel and the
environment ( CDC 1997).
The laboratory is not necessarily
separated from the general traffic
patterns in the building

7. BSL 1 (CONT)
 Work is generally conducted on open bench
tops using standard microbiological
practices.
 Special containment equipment or facility
designs is not required nor generally used.
 Insect and rodent pest control program
should be in effect
 Laboratory personnel have specific training
in the procedures conducted in the
laboratory and are supervised by a scientist
with general training in microbiology.

8. BSL 1(CONT)
The following standard special practices,
safety equipment, and facilities apply to agents
assigned to all Biosafety levels.
 Standard Microbiological Practices
 Access to the laboratory are limited or
restricted when experiments or work with
specimens and cultures are in progress
 After handling viable materials and animals
hands must be washed after removing gloves
and before leaving the laboratory.

9. STANDARD
MICROBIOLOGICAL
PRACTICES
Eating, drinking, smoking, handling contact
lenses, and applying cosmetics are not
permitted in the work areas where there is
reasonable likelihood of exposure to potentially
infectious materials. Persons who wear contact
lenses in laboratories should also wear goggles
or a face shield. Food is stored outside the work
area in cabinets or refrigerators designated and
used for this purpose only.
 Mouth pipetting is prohibited; mechanical
pipetting devices are used.

10. STANDARD
MICROBIOLOGICAL
PRACTICES
All procedures are performed carefully to minimize the
creation of splashes or aerosols.
Work surfaces are decontaminated at least once a day and
after any spill of viable material.
All cultures, stocks, and other regulated wastes are
decontaminated before disposal by an approved
decontamination method.
Materials to be decontaminated outside of the immediate
laboratory are to be placed in a durable, leak-proof container
and closed for transport from the laboratory.
Materials to be decontaminated at off-site from the laboratory
are packaged in accordance with applicable local, state, and
federal regulations, before removal from the facility.

11. SAFETY EQUIPMENT
Special containment devices or equipment such as a
biological safety cabinet are generally not required for
manipulations of agents assigned to Biosafety Level 1.
It is recommended that laboratory coats, gowns, or uniforms
be worn to prevent contamination or soiling of street clothes.
Gloves should be worn if the skin on the hands is broken or if
a rash exists.

12. SAFETY EQUIPMENT
Protective eyewear should be worn for anticipated splashes of
microorganisms or other hazardous materials to the face.

13. LABORATORY
FACILITIES
Each laboratory contains a sink for hand
washing.
The laboratory is designed so that it can be
easily cleaned. Rugs in laboratories are not
appropriate, and should not be used
because proper decontamination following a
spill extremely difficult to achieve.
Bench tops are impervious to water and
resistant to acids, alkalis, organic solvents,
and moderate heat.

14. LABORATORY
FACILITIES (CONT)
Laboratory furniture is sturdy. Spaces
between benches, cabinets, and equipment
are accessible for cleaning.
If the laboratory has windows that open,
they are fitted with fly screens.

15. BIOSAFETY LEVEL
2

16. BIOSAFETY CONTAINMENT LEVEL 2
Risk Group 2 infectious agents are pathogens
that can cause human or animal disease but,
under normal circumstances, are unlikely to be a
serious hazard to laboratory workers, the
community, livestock, or the environment
Level 2 infections are not considered to be a
serious hazard. They are a moderate individual
risk and limited community risk.

17. BIOSAFETY CONTAINMENT
LEVEL 2
Laboratory exposures rarely cause infection
leading to serious disease; effective treatment and
preventive measures are available and the risk of
spread is limited.
Examples of infectious agents in this risk level are
E. coli, California encephalitis viruses, many
influenza viruses, some fungi like ringworm;
California encephalitis viruses; human herpes
simplex viruses; many influenza viruses;
Transmissible Gastro-enteritis of swine; Mouse
Hepatitis Virus; and a few parasites.

18. PROCEDURES REQUIRED TO
CONTAIN RISKGROUP 2 ORGANISMS
 The laboratory should be separated from all other
activities.
A biohazard sign should be present and visible.
All the surfaces in the laboratory should be readily
cleanable and impervious.
An auto clave should be present.
A certified HEPA filtered class 1 or 2 biological cabinet
should be available for the manipulation of organisms.
Gloves and a laboratory coat should be worn at all times
in the laboratory.

19. BIOSAFETY LEVEL
3

20. BIOSAFETY CONTAINMENT
LEVEL 3
Risk Group 3 infectious agents are pathogens that usually
cause serious human or animal disease, or which can
result in serious economic consequences, but do not
ordinarily spread by casual contact from one individual to
another (high individual risk, low community risk), or that
can be treated by antimicrobial or antiparasitic agents.
 Risk Group 3 pathogens include bacteria such as
anthrax, Q Fever, tuberculosis, and viruses such as hanta
viruses, Human immunodeficiency viruses (HIV - all
isolates), eastern and western equine encephalitis viruses.

21. PROCEDURES REQUIRED TO
CONTAIN RISKGROUP 3 ORGANISMS
Laboratory staff must be fully trained in the handling of
pathogenic and other hazardous material, in the use of
safety equipment, disposal techniques, handling of
contaminated waste, and emergency response.
Standard operating procedures must be pasted in visible
spot.
Equipment must include an autoclave and a certified
HEPA filtered class II biological safety cabinet
All activities involving infectious materials to be conducted
in biological safety cabinets or other appropriate
combinations of personal protective and physical
containment devices.

22. PROCEDURES REQUIRED TO
CONTAIN RISKGROUP 3
ORGANISMS
Personal protective equipment should include
head covers and dedicated footwear, gloves,
appropriate respiratory protection
Specialized design and construction of
laboratories, with controlled access double door
entry and body shower.
Air pressure must be negative at all times, no
recirculation, HEPA filtration must be used.

23. BIOSAFETY
LEVEL 4
The Galveston
National
LaboratoryBSL-4
(P4) lab on the
Campus of
theUniversity of
Texas Medical
Branch

24. BSL- 4
Designed for use with:
Highly toxic/infectious agents
Agents that are at a very high risk for forming infectious aerosols
Life threatening agents
Ex. Ebola, Marburg, Lassa

25. SPECIAL PRACTICES
 Only those that work in the BSL-4 lab will be allowed entry
 Immuno-compromised individuals such as children and pregnant
women are not allowed to enter
 A logbook must be signed each time one comes in or out of the lab
with the date and time

26. SPECIAL PRACTICES
 The doors must have a universal hazard label with….
• Description of the infectious agent (s)
• Responsible director
• PPE required such as respirators and any immunization
requirements
 All lab personnel are trained specially for BSL-4 by lab
director

27. SPECIAL PRACTICES
 Lab personnel must be vaccinated for the agents…
• Present in the lab
• That have potential to be in the lab
 Baseline serum samples are collected and stored for serological
surveillance
• Makes sure the personnel have sufficient antibodies for the pathogens they are
working with
 Biosafety manual is adopted for that lab

28. SPECIAL PRACTICES
 Entering the lab
• Personnel must remove personal clothing in an outer
room and leave it
• All clothing and PPE is provided and expected to be
worn
 Exiting the lab
• Personnel remove lab clothing in the inner room
after taking a decontaminating shower
 The lab clothing is autoclaved then cleaned
• They exit through the outer room where their
personal clothes are

29. SPECIAL PRACTICES Supplies/materials are brought into the lab by a
double door autoclave that….
• is secured so that materials can be removed from
the interior door
• is decontaminated after each use
 Plastic is substituted for glass when possible
• Eliminate sharps such as capillaries, scalpels,
needles and syringes
• Broken glass should be handled with a dust pan and
broom
• Non-disposable and disposable sharps collected in a
hard container to be transported to an autoclave for
decontamination; disposables are thrown out

30. SPECIAL PRACTICES
 Any viable materials transferred from the class III Biological Safety
Cabinet or the lab itself must be put in a primary container and
then in a non- breakable secondary container
• It is then removed by an airlock
• Pressurizes the chamber so that minimal air is emitted when each door opens
one at a time
• Can also decontaminate the containers

31. SPECIAL PRACTICES
 Lab equipment is decontaminated after use,
after spills or before any repairs
 Any material not related to the project is not
permitted in the laboratory
 A system is set up for emergencies such as
exposure to infectious agents and also
employee absenteeism on safety
• Quarantine
• Medical care for those with lab acquired illness
• Records are kept for all accidents

32. SAFETY EQUIPMENT
(PRIMARY BARRIERS)
 Class III biological safety cabinets for handling
infectious material or….
 Class II biological safety cabinet in conjunction
with a positive pressure suit ventilated by a life
support system
 BSC must be validated on a 12 month period
 The PPE required is a…
• one-piece jumpsuit
• gloves
• closed toe shoes
• All will be removed in the inner room after showering

33. LABORATORY FACILITY
(SECONDARY BARRIER) BSL-4 labs are housed in separate buildings or isolated
within a building with lower BSL labs
 Outer and inner change rooms
 Walls, floors and ceilings have sealed internal shells
• keep pests out
• Liquid and heat stabile for decontaminating purposes
 Drains have chemical disinfectant traps attached to liquid
waste decontamination system
• All waste must be decontaminated including shower waste
 HEPA filters in vents for sterile air
• Filter air exhaust from the class III and II biological safety
cabinets and
• From ventilated positive pressure suits worn in class II safety
cabinets

34. LABORATORY FACILITY
(SECONDARY BARRIER)
 Windows are break resistant
 A dunk tank, fumigation chamber or airlock is
provided for safe passage of materials
outside/inside lab that cannot be
decontaminated in the autoclave
 A non-recirculation ventilation system is
provided with directional inward airflow
• Air flows from the outside of the lab to the inside but not vice versa

35. VIDEO ABOUT BIOSAFETY
LEVELS

36. REFERENCES
http://www.cdc.gov/OD/OHS/biosfty/bmbl/section3.htm